Analytical Chemistry Laboratory 2

Spectrophotometric Analysis of a Two Component Mixture

Mr. *****

Department of Chemical Engineering and Chemistry, Mapua Institute of Technology

Chromium and Cobalt ions both absorb visible light though their absorbance maxima are
fairly well separated. By measuring the absorbance at two different wavelengths of a solution
containing both ions, it is possible to simultaneously determine the concentration of each ion in
the solution. An unknown solution containing these species was analyzed using a
spectrophotometer. The concentration of cobalt in the mixture calculated using Beer’s law was
0.0131 M, while chromium was 0.0185 M.

is a proportionality constant called the molar

Objective absorptivity. ε has units of M-1 cm-1 when b

The purpose of this experiment is to
introduce the proper operation of a
spectrophotometer and how its use relates to
chemical analysis. This will be achieved by
recording absorbance measurements of a
two component mixtures, and calculating its
concentrations using Beer’s Law.
Discussion
Overview
When a beam of parallel radiation
passes through a layer of solution having a
thickness, b (cm) and a concentration, c
(moles/liter) of an absorbing species, a
consequence of interactions of the photons
and the absorbing particles is attenuation of
the beam. The transmittance (T) of the
solution is the fraction of the incident
radiation transmitted by the solution. The
absorbance (A) of a solution is defined as
the negative log of the transmittance (T) of
the solution. The absorbance is directly
proportional to the path length through the
solution and the concentration of the
absorbing species. That is, A= εbc where ε
and c are expressed in cm and moles per
liter respectively. This relationship between
absorbance (A) and εbc is known as Beer's
Law. Beer's Law is successful in describing
the absorption behavior of dilute solutions
only. At high concentrations, the average
distance between the species responsible for
absorption is diminished to the point where
each effects the charge distribution of its
neighbors. This interaction, in turn, can alter
their ability to absorb a given wavelength of
radiation. Because the extent of interaction
depends upon concentration, the occurrence
of this phenomenon causes deviations from
the linear relationship between absorbance
and concentration. A similar effect is
sometimes observed in solutions containing
high concentrations of electrolytes. The
proximity of ions (in addition to other
factors such as temperature) alters the molar
absorptivity of the absorbing species.
Methodology
The spectrophotometer was set to
the 575 nm chromium maximum. The
absorbance reading of a 0.02 M solution of
cobalt nitrate at this wavelength was taken,
including the absorbance of the unknown
chromium-cobalt mixture. The wavelength
of the spectrophotometer was adjusted again
to about 510 nm and the maximum was
located for cobalt absorption using a 0.06 M
solution of cobalt nitrate. After the
maximum was located, the wavelength value
was recorded. The absorbance of the 0.06 M
cobalt nitrate solution, the absorbance of the
0.04 M chromium nitrate solution, and the
absorbance of the unknown chromium-
cobalt mixture was read.
Instrumentation
A spectrophotometer measures the
amount of radiant energy absorbed by a
species. It consists of a source (incandescent
filament) that emits a continuous range of
wavelengths. The radiant energy is focused
by optics which includes a prism or grating;
these define a beam of radiant energy of a
specific wavelength range. The detector
determines the intensity of the radiation
before and after the beam is passed through
the sample in a cell:
Interpretation
In this experiment, a two component
solution will be studied. The absorbing
species are chromium ion and cobalt ion,
both of which absorb strongly in the visible
spectrum. The following data was obtained
from the experiment:
0.06 M Cr soln. 0.02 M Co soln. Sample
AλCr 0.8017 0.0158 0.2575
AλCo 0.314 0.0984 0.1661
In simultaneous determinations of
two species it is necessary to generate two
equations in order to determine the two
unknown concentrations. In a
spectrophotometric analysis these equations
can be developed from the Beer-Lambert
Law. Absorbance = A = e b C
To solve the simultaneous equations
that are generated when applying Beer's law
to mixtures of absorbing species, one must
know the molar absorptivity of each
component at each wavelength. The molar
absorbtivity of each compound at a given
wavelength can be determined from the
relationship between A and c in Beer’s law.
Beer's Law requires the use of
monochromatic radiation and it is under this
restraint that the linear dependence of
absorption and concentration occurs. If two
or more species in a sample absorb at a
specific wavelength, the instrument cannot
distinguish between the individual species; it
can only determine the total absorbance of
the sample. In the mixture of the two
species, Cr and Co, absorb at the same
wavelength the total absorbance at that
wavelength is:
Atotal = ACr + ACo
and,
Atotal = e1bCCr + e2bCCo
The most convenient way to
construct two equations is to measure the
total absorbance of the solution at two
different wavelengths, 510 nm and 575 nm.
Solving simultaneously: CCo = 0.0131 M and
CCr = 0.0185 M.
Conclusion
 The spectrophotometer was used in
the analysis of the two components in the
mixture which is cobalt and chromium. The
calculation basically focuses on one formula
– the Beer’s law. Beer's law states that
absorbance of electromagnetic radiation is
directly proportional to concentration: if
there is more than one absorbing species in
solution, the total absorbance is the sum of
the individual absorbance of all the
absorbing species, provided there is no
interaction among the various species. By
measuring the absorbance at different
wavelengths (510 and 575 nm), different
absorbing components could theoretically be
measured by this technique. Hence
calculating concentration from absorbance
using Beer’s law: CCo = 0.0131 M and CCr =
0.0185 M

References

1. Christian, G.D., Analytical Chemistry, 6th

edition. New Jersey. John Wiley, 2004.

2. Englis, D. T. and D. A. Skoog, Ind. Eng.

Chem. Anal. Ed., l5, 748.